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Porcine TNF-alpha Quantikine ELISA Kit Best Seller

R&D Systems, part of Bio-Techne | Catalog # PTA00

R&D Systems, part of Bio-Techne
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PTA00

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

5 pg/mL

Assay Range

23.4-1500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Porcine TNF-alpha Quantikine ELISA Kit

The Quantikine Porcine TNF-alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure porcine TNF-alpha in cell culture supernates, serum, and plasma samples. It contains E. coli-expressed recombinant porcine TNF-alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately measure recombinant porcine TNF-alpha. Results obtained using natural porcine TNF-alpha show dose response curves that are parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural porcine TNF-alpha.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Porcine

Specificity

Natural and recombinant porcine TNF-alpha

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 79 260 646 88 279 679
Standard Deviation 4.9 9.2 27 8.8 24 44
CV% 6.2 3.5 4.2 10.0 8.6 6.5

EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 104 358 880 106 348 866
Standard Deviation 7.2 13 36 9.7 32 72
CV% 6.9 3.6 4.1 9.2 9.2 8.3

Recovery for Porcine TNF-alpha Quantikine ELISA Kit

The recovery of porcine TNF-alpha spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 99 91-105
EDTA Plasma (n=6) 98 83-111
Heparin Plasma (n=4) 91 82-99
Serum (n=6) 105 91-120

Linearity

To assess the linearity of the assay, six samples containing and/or spiked with various concentrations of porcine TNF-alpha in each matrix were diluted with the appropriate Calibrator Diluent and then assayed.

Porcine TNF-alpha ELISA Linearity

Scientific Data Images for Porcine TNF-alpha Quantikine ELISA Kit

Porcine TNF-alpha ELISA Cell Culture Supernate Standard Curve

Porcine TNF-alpha ELISA Cell Culture Supernate Standard Curve

Porcine TNF-alpha ELISA Serum/Plasma Standard Curve

Porcine TNF-alpha ELISA Serum/Plasma Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TNF-alpha

Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71% - 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness.

Cleavage of membrane bound TNF-α by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.

Long Name

Tumor Necrosis Factor alpha

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Gene Symbol

TNF

Additional TNF-alpha Products

Product Documents for Porcine TNF-alpha Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Porcine TNF-alpha Quantikine ELISA Kit

For research use only

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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.